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Ribosomes that synthesize proteins are among the most central and evolutionarily conserved organelles. Given the key role of proteins in cellular functions, prokaryotic and eukaryotic pathogens have evolved potent toxins to inhibit ribosomal functions and weaken their host. Many of these ribotoxin-producing pathogens are associated with food. For example, food can be contaminated with bacterial pathogens that produce the ribotoxin Shiga toxin, but also with the fungal ribotoxin deoxynivalenol. Shiga toxin cleaves ribosomal RNA, while deoxynivalenol binds to and inhibits the peptidyl transferase center. Despite their distinct modes of action, both groups of ribotoxins hinder protein translation, but also trigger other comparable toxic effects, which depend or not on the activation of the ribotoxic stress response. Ribotoxic stress response-dependent effects include inflammation and apoptosis, whereas ribotoxic stress response-independent effects include endoplasmic reticulum stress, oxidative stress, and autophagy. For other effects, such as cell cycle arrest and cytoskeleton modulation, the involvement of the ribotoxic stress response is still controversial. Ribotoxins affect one organelle yet induce multiple toxic effects with multiple consequences for the cell. The ribosome can therefore be considered as the cellular "Achilles heel" targeted by food borne ribotoxins. Considering the high toxicity of ribotoxins, they pose a substantial health risk, as humans are highly susceptible to widespread exposure to these toxins through contaminated food sources. [ABSTRACT FROM AUTHOR]

Maize is a worldwide crop yet can be associated with mycotoxigenic fungi, much investigated in humid tropical and cooler, wet temperate regions. However, in hot, arid/semi-arid regions data on their occurrence are poor. In this paper, we focused on interactions between maize and Fusarium fungal species in Tunisia, which has a Mediterranean climate, with hot, dry summers and milder, damper winters. Maize kernels, stalks, and roots were sampled from 19 agricultural fields and 56, 72, and 88 % of samples, respectively, yielded Fusarium isolates. Based on molecular identifications, these were mainly F. verticillioides (67 %), and other species of Fusarium fujikuroi species complex and members of Fusarium incarnatum-equiseti- , oxysporum- , burgessii- , solani- and concolor species complexes. In addition, five isolates were identified as Clonostachys rosea. Fusarium verticilloides and Fusarium proliferatum , that produce fumonisins, suspected carcinogenic compounds, were isolated from all kinds of samples, whereas the other species were isolated only from root and stems. Fumonisin B 1 was higher in kernels than in silage, while deoxynivalenol, potent protein synthesis inhibiting compound, was detected (at low levels) in grains and silage. A subset of selected strains, representative of all species identified, was also used to evaluate their ability to produce mycotoxins. Fusarium verticillioides , F usarium proliferatum and Fusarium nygamai produced high levels of fumonisin B 1 in vitro , as well as beauvericin and enniatins. These findings confirm that, even in hot arid regions, which generally do not favour fungal growth, mycotoxin-producing fungi can be reason of concern for human and animal health. • We studied maize– Fusarium associations and their mycotoxins in Tunisia. • Tunisia, with a Mediterranean climate, is a paradigm of the northern African region. • Isolates of six Fusarium species complexes were obtained from maize. • Fusarium verticillioides was the most prevalent (on all parts of the plants). • Fumonisin production, in both kernels and silage, was not dependent on fungal growth. [ABSTRACT FROM AUTHOR]

Background: The aim of this study was to investigate the role of necroptosis in deoxynivalenol (DON)-induced liver injury and inflammation in weaned piglets. Methods: In Exp. 1, 12 weaned piglets were divided into 2 groups including pigs fed basal diet and pigs fed diet contaminated with 4 mg/kg DON for 21 d. In Exp. 2, 12 weaned piglets were divided into 2 groups including control piglets and piglets given a gavage of 2 mg/kg body weight (BW) DON. In Exp. 3, 24 weaned piglets were used in a 2 × 2 factorial design and the main factors including necrostatin-1 (Nec-1) (DMSO or 0.5 mg/kg BW Nec-1) and DON challenge (saline or 2 mg/kg BW DON gavage). On 21 d in Exp. 1, or at 6 h post DON gavage in Exp. 2 and 3, pigs were killed for blood samples and liver tissues. Liver histology, blood biochemical indicators, and liver inflammation and necroptosis signals were tested. Results: Dietary or oral gavage with DON caused liver morphological damage in piglets. Dietary DON led to hepatocyte damage indicated by increased aspartate transaminase (AST) activity and AST/alanine aminotransferase (ALT) ratio, and DON gavage also caused hepatocyte damage and cholestasis indicated by increased AST and alkaline phosphatase (AKP) activities. Dietary DON caused liver necroptosis indicated by increased protein abundance of total receptor interacting protein kinase 3 (t-RIP3) and total mixed lineage kinase domain-like protein (t-MLKL). Moreover, DON gavage increased mRNA expression of interleukin (IL)-6 and IL-1β in liver. DON gavage also induced liver necroptosis demonstrated by increased protein abundance of t-RIP3, phosphorylated-RIP3 (p-RIP3), t-MLKL and p-MLKL. However, pretreatment with Nec-1, a specific inhibitor of necroptosis, inhibited liver necroptosis indicated by decreased protein expression of t-RIP3, p-RIP3, t-MLKL and p-MLKL. Nec-1 pretreatment reduced liver morphological damage after DON gavage. Pretreatment with Nec-1 also attenuated liver damage induced by DON indicated by decreased activities of AST and AKP. Furthermore, Nec-1 pretreatment inhibited liver mRNA expression of IL-6 and IL-1β after DON challenge. Conclusions: Our data demonstrate for the first time that necroptosis contributes to DON-induced liver injury and inflammation in piglets. [ABSTRACT FROM AUTHOR]

The effect of mycotoxin exposure on follicular fluid composition and reproductive outcomes in women undergoing in vitro fertilisation (IVF) was investigated in this study. Twenty-five patients were included, and follicular fluid and serum samples were analysed for various mycotoxins. Principal observations:1. Mycotoxin presence: All examined mycotoxins were detected in follicular fluid. Follicular fluid (ff) levels: Deoxynivalenol (DON), alfa-Zearalenol (α-ZOL), Zearalenone (ZEN), and total aflatoxin (AFs) were significantly higher in follicular fluid than in serum. 2. Follicular fluid and reproductive outcomes: A positive correlation was observed between the ratio of oocytes to total follicles and the follicular Fumonisin B1 (FB1) levels. Multiple linear regression analysis revealed a significant relationship between DON and T-2/HT-2 toxins (T2/HT2) levels in the follicular fluid. 3. Hormone levels: Follicular 17-beta estradiol (E2) and progesterone (P4) levels were higher than the serum levels. Follicular P4 correlated with serum P4 and Anti-Müllerian hormone (AMH) levels. In contrast, follicular E2 did not correlate with plasma E2 levels. 4. Mycotoxin–hormone interactions: A positive correlation was observed between follicular P4 and T2/HT2 toxin levels, whereas a negative correlation was found between ffE2 and ffT2/HT2, and a positive correlation was found between ZEN and E2. Conclusion: This study elucidated the presence of various mycotoxins in the follicular fluid and their potential influence on reproductive outcomes. Further research is warranted to clarify the specific mechanisms underlying these effects and develop strategies for detecting mycotoxin exposure in women undergoing IVF. [ABSTRACT FROM AUTHOR]

Gold-grafted melamine sponge (AuSp) was fabricated and applied as surface-enhanced Raman spectroscopy (SERS) substrate for enzyme-linked immunoassay of 2 typical mycotoxins, zearalenone (ZEN) and T-2 toxin. The gold nanoparticles were in situ grown on the sponge, and the product showed good effect in enhancing Raman signal and good stability. COMSOL and Gaussian simulations were used to analyze the enhancing effect. Some of the experimental conditions that may affect the determination results were studied in order to meet the maximum response to mycotoxins. At optimized conditions, the limits of detection of the method were 1 μg·kg−1 and 0.05 μg·kg−1 for ZEN and T-2 toxin, with linear ranges of 5–100 μg·kg−1 and 0.1–20 μg·kg−1, respectively, and relative standard deviations lower than 7.0%. The established method was used to determine ZEN and T-2 toxin in six kinds of cereal samples, the recoveries were 85–120%, and relative errors were less than 13.0%. These results indicated the accuracy of the established method. After 1-year storage, signal strength of AuSp for mycotoxins determination decreased by less than 10%, showing stability over a long time. Our work provided a new method for SERS determination of mycotoxins, which reduces the complexity of substrate preparation and the sample consumption compared with traditional methods, and thus has potential for on-line SERS determination. [ABSTRACT FROM AUTHOR]

Fusarium graminearum is a fungal species affecting the quality and safety of malting barley grains, one of the most important cereals worldwide. Fungal growth and mycotoxin production vary among growing seasons and sowing locations, mainly due to weather conditions. This work aimed to assess the incidence of F. graminearum and the contamination with Deoxynivalenol (DON) in 40 barley grain samples from different Buenos Aires, Argentina localities during the 2017 and 2018 growing seasons. F. graminearum was identified in 80% of the samples. It was isolated in eight of eleven localities in the first and ten in the second growing seasons, with a similar maximum incidence (20% and 17%, respectively). On the other hand, all samples were contaminated with DON, and 75% exceeded the maximum limits established by The European Union (EC 1126/2007). The level of DON contamination was significantly higher in the second growing season, which was rainier and had a higher mean temperature (an average of 2.5 ppm in 2017 and 3.75 ppm in 2018). The results obtained in the present study show the need to establish regulations in Argentina on maximum limits of Fusarium mycotoxins in barley. [ABSTRACT FROM AUTHOR]

Mycotoxin contamination in agricultural goods is a major global problem due to its negative impact on human and animal health. The principal mycotoxin producers are fungal species from the genera Fusarium, Aspergillus, Alternaria, and Penicillium. The toxigenic fungal species produce the mycotoxins as secondary metabolites when they invade agricultural commodities during crop cultivation in the field (preharvest) or after harvesting or during transport and storage. This study was designed to investigate the levels of Fusarium mycotoxins, viz., fumonisin (FUM), zearalenone (ZEN), and deoxynivalenol (DON) in cereal grain samples collected from Muscat, Sultanate of Oman during 2023-24. A total of 90 cereal grain (wheat, corn, rice, barley) samples from local markets at Muscat, the Plant Quarantine Department, Oman, and Oman Flour Mills Company were analyzed using competitive enzyme immunoassay kits. Furthermore, Fusarium spp. associated with the contaminated grain samples were isolated, and their mycotoxin-producing potential was assessed. The results indicated that FUM, ZEN, and DON levels were below the detection limit (LOD) in 81%, 97%, and 44% of the samples, respectively. Two out of fifteen corn samples and one out of thirty-seven wheat samples tested exceeded the maximum permissible limit for FUM and ZEN, respectively, as set by the European Commission. A total of 19 Fusarium spp. associated with the contaminated grain samples were isolated and identified through molecular techniques. Sixteen isolates of F. verticillioides, one isolate of F. thapsinum, and two new Fusarium species were identified based on nuclear ribosomal DNA internal transcribed spacer and elongation factor 1-alpha sequences. Two isolates of F. verticillioides (FQD-1 and FQD-20) produced FUM levels exceeding 2000 µg kg−1. The maximum ZEN concentration was observed in F. verticillioides FQD-20 (9.2 µg kg−1), followed by F. verticillioides FQD-2 (2.8 µg kg−1) and Fusarium sp. FOFMC-26 (2.5 µg kg−1). All tested Fusarium strains produced DON, with levels ranging from 25.6 to 213 µg kg−1, with F. thapsinum FQD-4 producing the highest level (213 µg kg−1). To our knowledge, this is the first report on the occurrence of Fusarium mycotoxins and mycotoxigenic Fusarium spp. in food commodities in Oman. [ABSTRACT FROM AUTHOR]

The contamination of food and feed by mycotoxins, particularly zearalenone (ZEA) and deoxynivalenol (DON), is a global issue. Prenatal exposure to ZEA and DON can result in congenital cardiac malformations in fetuses. Addressing the prevention and mitigation of embryonic cardiotoxicity caused by these toxins is crucial. Citrus limonoid nomilin (NOM) is an extract known for its pathological properties in various diseases. This study investigated the potential mechanism of NOM in mitigating cardiotoxicity caused by ZEA and DON co-exposure in a zebrafish model. The findings indicated that NOM pretreatment alleviated cardiac developmental toxicity induced by ZEA and DON and normalized the expression of key genes involved in heart development, including gata4, vmhc, nkx2.5, and sox9b. Co-exposure to NOM, ZEA, and DON enhanced SOD and catalase activity, increased glutathione levels, and reduced ROS and malondialdehyde production. Furthermore, NOM reduced cardiac oxidative damage by activating the Keap1/Nrf2 signaling pathway. In summary, this study offers new insights for preventive interventions against congenital heart disease caused by mycotoxin exposure. [ABSTRACT FROM AUTHOR]

(1) T-2 toxin synthesized by Fusarium oxysporum (F. oxysporum) can cause deterioration of dried fish and endanger human health. (2) The molecular mechanism by which antibacterial lipopeptides surfactin inhibited F. oxysporum growth and toxin production was elucidated by investigating the intracellular ROS production pathway and the subcellular distribution and transcriptional activity of the transcription factor Yap1 and its regulation of Tri5 gene in F. oxysporum. (3) Surfactin caused hyphal damage and nucleic acid and protein leakage; thus, the growth of F. oxysporum was disrupted. Surfactin's excessive accumulation of intracellular ROS triggered the translocation of transcription factor Yap1 into the nucleus, resulting in toxin cluster gene Tri5 expression inhibition, thereby blocking T-2 toxin synthesis. (4) This is a novel mechanism by which surfactin inhibits the growth and T-2 toxin synthesis of F. oxysporum from multiple aspects, including cell structural integrity and the ROS-Yap1 signaling pathway. (5) This study provides a theoretical basis for the application of surfactin in the antifungal control of aquatic dry products. [ABSTRACT FROM AUTHOR]

A QuEChERS (quick, easy, cheap, effective, rugged, and safe)-based multi-mycotoxin method was developed, analyzing 24 (17 free and 7 modified) Alternaria and Fusarium toxins in cereals via ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). A modified QuEChERS approach was optimized for sample preparation. Quantification was conducted using a combination of stable isotope dilution analysis (SIDA) for nine toxins and matrix-matched calibration for ten toxins. Quantification via a structurally similar internal standard was conducted for four analytes. Alternariol-9-sulfate (AOH-9-S) was measured qualitatively. Limits of detection (LODs) were between 0.004 µg/kg for enniatin A1 (ENN A1) and 3.16 µg/kg for nivalenol (NIV), while the limits of quantification were between 0.013 and 11.8 µg/kg, respectively. The method was successfully applied to analyze 136 cereals and cereal-based foods, including 28 cereal-based infant food products. The analyzed samples were frequently contaminated with Alternaria toxins, proving their ubiquitous occurrence. Interestingly, in many of those samples, some modified Alternaria toxins occurred, mainly alternariol-3-sulfate (AOH-3-S) and alternariol monomethyl ether-3-sulfate (AME-3-S), thus highlighting the importance of including modified mycotoxins in the routine analysis as they may significantly add to the total exposure of their parent toxins. Over 95% of the analyzed samples were contaminated with at least one toxin. Despite the general contamination, no maximum or indicative levels were exceeded. [ABSTRACT FROM AUTHOR]

[Objectives] Heterogeneous nuclear ribonucleoproteins (hnRNP) represent a large family of RNA-binding proteins that contribute to multiple aspects of RNA metabolism. In Fusarium graminearum, FGSG_07478 was identified as the homologous protein of annotated Fusarium hnRNP G. The objective of this study was to analyze the function of FGSG_07478 in growth and development, stress, mycotoxins production and pathogenicity of F.graminearum. [Methods] Based on homologous recombination and polyethylene glycol (PEG) mediated protoplast transformation, the FGSG_07478 gene knockout mutant strain ΔFGSG_07478 was obtained to investigate its function in mycelial growth, asexual and sexual reproduction, and stress resistance of F.graminearum. Pathogenicity of the mutant ΔFGSG_07478 was assessed by infection assays using wheat coleoptiles. Deoxynivalenol (DON) produced by ΔFGSG_07478 was quantified using the ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method, and the relative expression levels of seven TRI involved in DON biosynthesis were determined by RT-qPCR. [Results] Compared with the wild type strain PH-1, the growth rate of ΔFGSG_07478 was reduced by 16.2%, and the spore production decreased by 43.4%. In mating cultures, the ΔFGSG_07478 mutant produced less perithecia, verifying the weakened sexual reproduction. External environmental stress assays showed that ΔFGSG_07478 mutant became sensitive to H2O2 and SDS, but more resistant to Congo red. Although no significant difference in pathogenicity, DON biosynthesis and expression levels of TRI4, TRI5, TRI6, TRI8, TRI10, TRI12 and TRI101 were significantly reduced in ΔFGSG_07478 mutant compared with the wild type. [Conclusions] FGSG_07478 played an important role in fungal growth and development, sexual reproduction, responses to various environmental stresses and mycotoxins production in F.graminearum. [ABSTRACT FROM AUTHOR]

Mycotoxin cocontamination is a severe threat to health and economic security worldwide. The mycotoxins aflatoxin B1 (AFB1), zearalenone (ZEN), deoxynivalenol, T-2 toxin, fumonisin B1, and ochratoxin A are of particular concern due to their substantial toxicity. Lac-W is a laccase with the unique property of degrading these six mycotoxins in the absence of redox mediators. Nevertheless, their degradation rates are low. This work aims to improve the ability of Lac-W to degrade these six mycotoxins and to elucidate its detoxification mechanism. Including redox mediators increased the Lac-W degradation efficiency drastically, and completely degraded AFB1 and ZEN within one hour. Additionally, Lac-W-AS has good temperature, pH, and ions adaptability in ZEN degradation. Lac-W-AS reduced the ZEN toxicity because ZEN degradation products significantly restored the bioluminescence intensity of Vibrio fischeri. A Lac-W-AS-mediated oxidation product of ZEN was structurally characterized as 15-OH-ZEN by UHPLC-MS/MS. Linear sweep voltammetry showed that AS affected the potential of Lac-W and accelerated the oxidation of ZEN. Finally, the combination of mediators (acetosyringone and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonate)) improved the degradation rate of mycotoxins. This work highlights that the combination of Lac-W with mediators serves as a good candidate for degrading multi-mycotoxin contaminants in food and feedstuff. [ABSTRACT FROM AUTHOR]

This study investigated the effects of delayed harvesting, varying meteorological conditions, and barley variety on Fusarium spp. infection rates, nutritional composition, and mycotoxin contamination in barley grains. Field experiments were conducted from 2020 to 2022 and involved two barley varieties: 'Laureate' for malting and 'Luokė' for feed. The results indicated that the dominant Fusarium species isolated were F. avenaceum, F. culmorum, F. poae, F. sporotrichioides, F. tricinctum, and F. equiseti. These tended to increase in number with delayed harvest times and were more prevalent during harvest periods of higher precipitation (p < 0.05). Malting barley had higher starch and lower protein content compared to feed barley (p < 0.05). Delayed harvesting generally increased dry matter, crude fat, and crude ash contents while decreasing crude protein, zinc, and iron contents (p < 0.05). Mycotoxin analysis revealed significant differences under specific weather conditions. HT-2 toxin levels were higher under slightly warmer and wetter conditions during flowering, with harvest conditions similar to the long-term average. Zearalenone levels increased with dry, warm growing seasons followed by rainy harvests. Nivalenol and enniatin levels increased with rainy growing seasons and dry, warm harvests. Deoxynivalenol concentrations did not reach the limit of quantification throughout the study. No consistent trend was observed for higher contamination in any specific barley variety (p > 0.05). The strongest correlations between mycotoxins and nutritional value indicators were observed with less-studied mycotoxins, such as nivalenol and enniatins, which exhibited negative correlations with crude protein (p < 0.01), crude fat (p < 0.05), and zinc (p < 0.01), and positive correlations with crude ash (p < 0.05) and phosphorus (p < 0.01). [ABSTRACT FROM AUTHOR]

Summary: Food safety is crucial globally, particularly regarding mycotoxins in food. Mycotoxins, primarily found in cereals, can also contaminate animal products. In Mexico, staple foods like maize and animal products may contain fumonisins, mycotoxins from Fusarium fungi linked to various health issues. Type B fumonisins (FBs) and their hydrolysis products are particularly toxic to humans. This study analysed the most commonly consumed foods in the Mexican diet, selected based on data from Mexican government institutions. Samples were processed and dried at various temperatures according to the food type. To identify the most effective extraction method, each sample underwent testing under different extraction conditions. The presence of FB1, FB2, FB3, and their hydrolysed forms in the selected foods, as well as their relative concentrations, was determined using high‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry (HPLC‐MS‐QTOF). Therefore, this work aims to analyse the presence of fumonisins as contaminants in foods consumed by Mexicans, providing an analytical method for their inspection. [ABSTRACT FROM AUTHOR]

Fusarium verticillioides is both an endophyte and pathogen of maize. During growth on maize, the fungus often synthesizes the mycotoxins fumonisins, which have been linked to a variety of diseases, including cancer in some animals. How F. verticillioides responds to other fungi, such as Fusarium proliferatum , Aspergillus flavus , Aspergillus niger , and Penicillium oxalicum , that coinfect maize, has potential to impact mycotoxin synthesis and disease. We hypothesize that low molecular weight acids produced by these fungi play a role in communication between the fungi in planta/nature. To address this hypothesis, we exposed 48-h maize kernel cultures of F. verticillioides to oxalic acid, citric acid, fusaric acid, or kojic acid and then compared transcriptomes after 30 min and 6 h. Transcription of some genes were affected by multiple chemicals and others were affected by only one chemical. The most significant positive response was observed after exposure to fusaric acid which resulted in >2-fold upregulation of 225 genes, including genes involved in fusaric acid synthesis. Exposure of cultures to the other three chemicals increased expression of only 3–15 genes. The predicted function and frequent co-localization of three sets of genes support a role in protecting the fungus from the chemical or a role in catabolism. These unique transcriptional responses support our hypothesis that these chemicals can act as signaling molecules. Studies with gene deletion mutants will further indicate if the initial transcriptional response to the chemicals benefit F. verticillioides. • Low-molecular-weight fungal acids impact the Fusarium verticillioides transcriptome. • Fusaric acid impacted more genes than other acids. • Kojic acid impacted two gene clusters that occur widely in Fusarium. [ABSTRACT FROM AUTHOR]

ABSTRACT Mycotoxins, originating from contaminated raw materials or suboptimal feed storage, are a growing concern in tropical aquaculture. Common fungi such as Aspergillus spp. and Fusarium spp. produce mycotoxins including aflatoxin, fumonisin, deoxynivalenol and zearalenone. High doses or prolonged exposure (weeks) to low doses of these mycotoxins (< 20 μg/kg) can depress growth, immunity, and cause mortality. This study investigated poor growth and low survival in juvenile Asian seabass, Lates calcarifer fed two different diets (FM40 and ABS3) for 5 weeks. Analysis of these diets revealed high peroxide values, multiple mycotoxins and high histamine levels. Fish fed the FM40 diet, which was contaminated with aflatoxin B1 (13.2 μg/kg), aflatoxin B2 (1.9 μg/kg), deoxynivalenol (29.5 μg/kg), alternariol (2.2 μg/kg), elevated peroxide value (45.91 mEq/kg), and histamine (129.51 mg/kg) developed mild bile duct hyperplasia, depressed total serum proteins (50.40 ± 10.06 g/L), markedly elevated blood potassium (8.2 ± 0.18 mmol/L), and heavy iron deposits in splenic melanomacrophage centres (Perl's stain) indicative of increased haemolysis. The presence of multiple cytotoxic mycotoxins in FM40 diet could explain the increased haemolysis and elevated blood potassium. In contrast, fish fed the ABS3 diet, which had high histamine levels (210.05 mg/kg), exhibited protein‐losing nephropathy with multifocal fibrin plugs (Martius scarlet blue stain) indicating acute renal damage, and elevated blood calcium and phosphorus levels. Histamine is metabolised and excreted through the kidneys and known to induce renal arteriolar constriction, disrupt glomerular filtration barrier and increase permeability resulting in protein loss. This study shows that blood biochemistry and histopathology are useful diagnostic tools for assessing the impact of mycotoxins and histamines on fish health. [ABSTRACT FROM AUTHOR]

Fungal infection in agricultural grains is a global problem, particularly if it is accompanied by mycotoxin production. In this study, the degradation of mycotoxins by laccase and manganese peroxidase was investigated. Aspergillus flavus, Aspergillus fumigatus, and Fusarium graminearum were recorded in infected maize grains. Aflatoxin B1 (AF B1) was detected (from 3.38 to 2.60 ppm) on the infected samples by fungi compared to other detected aflatoxins. Trichothecene (T-2) toxin and deoxynivalenol (DON) were recorded with concentrations ranging from 0.464 to 0.184 ppm and 0.370 to 0.214 ppm, respectively. The addition of laccase and manganese peroxidase to the inoculated medium with A. flavus and F. graminearum individually degraded the produced AF B1, B2, G1, G2, T-2 toxin, and DON from 5.0, 1.33, 0.76, 0.61, 0.63, and 0.38 ppm to 2.77, 0.66, 0.37, 0.15, 0.45, and 0.38 ppm using laccase, to 3.08, 1.25, 0.61, 0.39, 0.55, and 0.36 ppm using manganese peroxidase. The computational technique (docking) demonstrated the laccase and manganese peroxidase activities on aflatoxin and DON degradation. Consequently, the results suggested that laccase (PDB ID: 1HFU) and manganese peroxidase (PDB ID: 1MNP) promise innovative activity toward aflatoxin degradation, while 1HFU has more effect than 1MNP on DON degradation. [ABSTRACT FROM AUTHOR]

Mycotoxins are a major threat to animal and human health, as well as to the global feed supply chain. Among them, aflatoxins, fumonisins, zearalenone, T-2 toxins, deoxynivalenol, and Alternaria toxins are the most common mycotoxins found in animal feed, with genotoxic, cytotoxic, carcinogenic, and mutagenic effects that concern the animal industry. The chronic negative effects of mycotoxins on animal health and production and the negative economic impact on the livestock industry make it crucial to develop and implement solutions to mitigate mycotoxins. In this review, we summarize the current knowledge of the mycotoxicosis effect in livestock animals as a result of their contaminated diet. In addition, we discuss the potential of five promising phytogenics (curcumin, silymarin, grape pomace, olive pomace, and orange peel extracts) with demonstrated positive effects on animal performance and health, to present them as potential anti-mycotoxin solutions. We describe the composition and the main promising characteristics of these bioactive compounds that can exert beneficial effects on animal health and performance, and how these phytogenic feed additives can help to alleviate mycotoxins' deleterious effects. [ABSTRACT FROM AUTHOR]

Maize is one of the most important crops cultivated worldwide, whose production can be affected by the presence of several pathogens. Fusarium verticillioides and Fusarium graminearum are the most predominant pathogens affecting maize ears. However, few studies have been focused on studying the interaction between both pathogens in field conditions. For this reason, the aim of the present work was to evaluate the interaction between F. graminearum and F. verticillioides in different genotypes of maize under field conditions. Field experiments were carried out during two growing seasons in Azul, Argentina, including 12 commercial hybrids of maize, which were inoculated with F. graminearum , F. verticillioides , and a mixture of both pathogens. Phenotypic traits (plant height, plant diameter, tiller and cob number, and radiation interception), disease evaluation, and mycotoxin contamination were analyzed. The results showed significant differences between genotypes in disease severity (DS) for both years. In general terms, higher values of DS were reported in 2020 (21.70% ± 0.40) than in 2021 (16.50% ± 0.20). Different climatic conditions registered along the assay, especially precipitations and relative humidity, could be responsible for the differences observed over the years. Moreover, no significant correlations were found regarding DS and mycotoxin contamination for each genotype. For these reasons, an automatic correspondence between DS and mycotoxin contamination could lead to wrong agronomic decisions. The present study points out novel information regarding plant–pathogen interaction (maize- F. verticillioides/F. graminearum) under field conditions that could be useful for future maize breeding programmes. [ABSTRACT FROM AUTHOR]

The present study revealed the consequences of the interaction of a widely used bioinsecticide and endophyte Metarhizium anisopliae with the hazardous mycotoxin zearalenone (ZEN) as a pure substance and with ZEN as a native component of a crude Fusarium extract. In the environment, microorganisms encounter a mixture of metabolites secreted by other organisms living in the same area, not single substances. The obtained results suggest that M. anisopliae, exposed to a variety of active substances produced by Fusarium graminearum, is able to eliminate ZEN. Within 14 days, M. anisopliae biotransformed 90.8% and 85.8% of ZEN as a pure substance and ZEN as a native component of the F. graminearum extract from Rice Medium (E-Fg-RM), respectively, through reduction predominantly to α-epimers of zearalenols and zearalanols, considered more estrogenic than ZEN, which can raise concerns. Compared to pure ZEN, E-Fg-RM significantly affected the production of Metarhizium secondary metabolites by increasing the destruxins amount by approximately 20–25% and reducing the swainsonine content by 96.2%. All these findings provide a possible picture of the interaction of M. anisopliae with ZEN in the wild, mainly as a result of the use of crude extract from Fusarium, which contained a mixture of different metabolites. [ABSTRACT FROM AUTHOR]

Fusarium graminearum is the main pathogenic fungus causing wheat head blight. It not only causes severe crop yield reduction, but also produces fungal toxins such as deoxynivalenol (DON) and 15-acetyl deoxynivalenol (15-ADON), which are dangerous food pollutants to people and livestock. For the genetic functional study of Fusarium graminearum, it is often necessary to culture mycelium on LTB medium in the laboratory to investigate its regulatory effect on DON and 15-ADON synthesis. However, the detection of DON and 15-ADON in the mycelium is focused on extracellular, with less focus on the detection of intracellular. Simultaneous detection of intracellular and extracellular DON and 15-ADON can provide more in-depth information of fungal toxin production and secretion functions. In the study, a method based on gas chromatography-mass spectrometry (GC-MS) for simultaneous detection of DON and 15-ADON was developed. The solvents for extracting DON and 15-ADON can be completely separated and the calibration curves of each extract show good linear relationship within a certain range. The mass spectrometric characteristic scan shows high ionization efficiency for both DON and 15-ADON. The retention time of DON and 15-ADON in chromatogram are 6.98 and 7.68 min, respectively. Qualitative analysis of DON was performed using m/z 295, 235, and 193 under single ion monitor (SIM) mode, while m/z 392, 235, and 193 were used for 15-ADON. The highest intensity ion m/z 235 was selected for the quantification of DON, and m/z 193 for the quantification of 15-ADON. This method was used to detect the intracellular and extracellular levels of DON and 15-ADON in wild-type PH-1 and tri5 mutant with DON biosynthesis deficiency. The intracellular DON and 15-ADON levels in PH-1 are determined to be (149.13±9.15) μg/g and (1 833.31±185.33) μg/g, extracellular levels are (5 910.35± 468.23) μg/g and (45 222.12±2 726.81) μg/g, respectively. The study identified a mutant strain with the deletion of the first enzyme involved in the synthesis pathway of DON and 15-ADON，both DON and 15-ADON are not detected in the extracellular and intracellular of the tri5 mutant. Additionally, this research confirms that after 7 days of cultivation in LTB medium, the wild-type strain exhibits higher levels of 15-ADON in both intracellular and extracellular compared to DON, with extracellular levels of DON and 15-ADON being higher than intracellular levels. This method can provide a methodological reference for the functional study of toxin synthesis in Fusarium graminearum. [ABSTRACT FROM AUTHOR]

The widespread presence of Fusarium mycotoxins in animal feed is a global issue, not only for the health of livestock but also for ensure the safety of food as an end product. High concentrations of zearalenone (ZEN) and deoxynivalenol (DON) have been detected in the diets of Japanese Black (JB) and Holstein Friesian (HF) breeding herds. Consequently, we monitored serum biochemical parameters over a long time in both herds, focusing on anti-Müllerian hormone (AMH) levels and acute-phase inflammation. Additionally, urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) and progesterone levels were measured in the HF herd. The JB herd, a ZEN-dominant model with low DON contamination, demonstrated ZEN levels that exceeded the Japanese limit in the purchased total mixed rations (TMR). Conversely, the HF herd, which primary consumes DON-dominant feed with low ZEN contamination, had high DON levels in the dent corn silage. Specifically, the JB herd's TMR contained 1.79 mg/kg ZEN and 0.58 mg/kg DON, whereas the HF herd's silage had 15.3 mg/kg DON (dried sample) and 0.1 mg/kg ZEN. Enzyme-linked immunoassay were used to measure urinary ZEN-DON levels following confirmation through liquid chromatography-tandem mass spectrometry. Urinary ZEN-DON levels measured were significantly correlated (p < 0.05, r > 0.6) in both herds. In the HF herd, AMH levels increased (p = 0.01) and serum amyloid A (SAA) levels decreased (p = 0.02) when contaminated and at the end of the monitoring period. Additionally, urinary ZEN and DON levels were significantly correlated with SAA levels (ZEN: p = 0.00, r = 0.46; DON: p = 0.03, r = 0.33), with an increase in ZEN and DON levels resulting in higher SAA levels. The JB herd showed no significant differences. Additionally, in the HF herd, 8-OHdG/Cre levels increased significantly during major contamination periods (p < 0.05). Clinical data from the HF herd indicated an increase in mastitis cases and treatment rates during periods of major contamination. Abortion rates in the HF herd decreased from 22.9% (before monitoring) to 8.9% (during the high contamination period) and finally to 1% (at the end of the monitoring period), with corresponding increases in progesterone levels. ZEN-DON contamination adversely affects breeding cattle's productivity, reproductive performance, and health. Therefore, monitoring urinary ZEN-DON is valuable for detecting contaminants and ensuring the safety of food products. [ABSTRACT FROM AUTHOR]

Mycotoxins have the potential to increase the risk of airway or intestinal infection due to their effects on epithelial integrity and function. The bacterium Streptococcus suis (S. suis) is often carried in pigs and can cause outbreaks of invasive disease, leading to sepsis and meningitis in postweaning piglets. In this study, we tested the effect of two Fusarium mycotoxins (deoxynivalenol (DON) and T-2) on the integrity of the intestinal epithelium and their interaction with S. suis. Porcine ileal organoids were exposed to DON and T-2 individually or in combination and co-cultured with or without S. suis. Both DON and T-2 were toxic for ileal organoid monolayers at a concentration of 1 µM but not S. suis, even at a higher concentration of 4 µM. To mimic sub-clinical exposures on farms, DON was tested at a concentration of 0.1 µM and T-2 at a concentration of 0.01 µM. The mycotoxins alone did not affect cell permeability, but in combination with S. suis there was an increase in epithelial permeability. Furthermore, DON and T-2 together decreased the transepithelial electrical resistance and increased bacterial translocation. [ABSTRACT FROM AUTHOR]

The quality of food is one of the emergent points worldwide. Many microorganisms produce toxins that are harmful for human and animal health. In particular, mycotoxins from Fusarium fungi are strictly controlled in cereals. Simple and robust biosensors are necessary for 'in field' control of the crops and processed products. Nucleic acid-based sensors (aptasensors) offer a new era of point-of-care devices with excellent stability and limits of detection for a variety of analytes. Here we report the development of a surface-enhanced Raman spectroscopy (SERS)-based aptasensor for the detection of T-2 and deoxynivalenol in wheat grains. The aptasensor was able to detect as low as 0.17% of pathogen fungi in the wheat grains. The portable devices, inexpensive SERS substrate, and short analysis time encourage further implementation of the aptasensors outside of highly equipped laboratories. [ABSTRACT FROM AUTHOR]

Context: The contamination of feedstuffs with multi-mycotoxins and their transfer to milk is an important concern because it can affect the health of both animals and humans. Aims: This study determined the nutrient composition and quantified the mycotoxins present in dairy feedstuffs and in cow's milk. Methods: Samples of feedstuffs and milk were collected from 11 dairy farms in northern Tunisia for physicochemical composition and mycotoxin analyses. The extraction of mycotoxins was performed by the QuEChERS procedure and the presence of 26 mycotoxins was checked using liquid or gas chromatography with tandem mass spectrometry (LC–MS/MS and GC–MS/MS). Key results: Nutrient contents differ among feedstuffs (P < 0.05). The pH values of oat, ryegrass and pulp silages varied from 4.6 to 5.3. Dry-matter (DM) content was highest for concentrate feeds and hay (90% and 86% DM respectively). Crude protein (CP) content was highest in concentrate feeds (15.8% DM). The neutral detergent fibre, acid detergent fibre and acid detergent lignin contents were highest in green grass, hay, and silage. Eight Fusarium mycotoxins, enniatins (ENA1, ENB, ENB1), beauvericine (BEA), HT-2 toxin (HT-2), deoxynivalenol (DON), 15-acetyldeoxynivalenol (15-ADON), Fumonisin B2 (FB2) and one Alternaria mycotoxin tentoxin (TENT) were detected at low concentrations. There was a significant (P < 0.05) difference in the levels of contamination by ENB between the samples of forages and concentrate feeds. The ENs (ENA1, ENB, ENB1) were the most frequently detected combination of mycotoxins. Milk analysis for fat, CP, lactose and dehydrated dry extract averaged 3.49%, 3.10%, 4.71%, and 8.77% respectively; urea concentration was of 34.01 mg/dL, milk pH was 6.5, and a somatic cell count of 1168.63 103 cells/mL was found. Most milk samples (82%) were contaminated with ENB. Conclusions: The levels of mycotoxin contamination in feedstuffs and milk were low and do not present a risk for dairy cattle and consumer health. Implications: Mycotoxins in feedstuffs and milk should be regularly analysed to make sure that quality and safety standards are met. The safety and nutritional quality of milk for human consumption depends on the quality of the feed sources consumed by dairy cattle. Our study has shown that mycotoxins, harmful toxins, detected in some feed sources (e.g. forage silages, hay and concentrates) could be transferred to milk. Strategies for efficient management of feed sources should be developed and the transferability of mycotoxins to milk should be controlled to ensure that the food supply chain is safe and free from harmful toxins. [ABSTRACT FROM AUTHOR]

Consumers are increasingly looking for healthier and sustainable diets. Plant-based diets rich in legumes satisfy this demand. Legumes contain protein, dietary fibers and starch. Technological processes can separate these fractions, which can be used as supplements, or as ingredients. Nonetheless, legumes are susceptible to fungal infection, causing a potential health concern, since some fungi can produce mycotoxins: toxic secondary metabolites. The aim of this work was to analyze the fate of mycotoxins during different stages of the production process of legume derived products from the raw materials to final products. An extraction followed by liquid chromatography-tandem mass spectrometry was used for the analysis, revealing the presence of enniatin B (ENN B), alternariol monomethyl ether (AME), deoxynivalenol, T2-toxin, nivalenol, fumonisin B1 and sterigmatocystin in raw materials, intermediate products and side streams. The alkaline solubilization steps, were effective in reducing ENN B; however, AME was found in one of the final products. [ABSTRACT FROM AUTHOR]

Fusarium Head Blight (FHB), predominantly caused by Fusarium species, is a devastating cereal disease worldwide. While considerable research has focused on Fusarium communities in grains, less attention has been given to residues and soil, the primary inoculum sources. Knowledge of Fusarium spp. diversity, dynamics, and mycotoxin accumulation in these substrates is crucial for assessing their contribution to wheat head infection and the complex interactions among Fusarium communities throughout the wheat cycle. We monitored six minimum-tillage wheat fields, with maize as the preceding crop, over two years. Soils, maize residues, and wheat grains were sampled at four stages. Fusarium composition was analyzed using a culture-dependent method, species-specific qPCR, and EF1α region metabarcoding sequencing, enabling species-level resolution. The Fusarium communities were primarily influenced by substrate type, accounting for 35.8% of variance, followed by sampling location (8.1%) and sampling stage (3.2%). Among the 32 identified species, F. poae and F. graminearum dominated grains, with mean relative abundances of 47% and 29%, respectively. Conversely, residues were mainly contaminated by F. graminearum, with a low presence of F. poae, as confirmed by species-specific qPCR. Notably, during periods of high FHB pressure, such as in 2021, F. graminearum was the dominant species in grains. However, in the following year, F. poae outcompeted F. graminearum, resulting in reduced disease pressure, consistent with the lower pathogenicity of F. poae. Source Tracker analysis indicated that residues were a more significant source of Fusarium contamination on wheat in 2021 compared to 2022, suggesting that F. graminearum in 2021 primarily originated from residues, whereas F. poae's sources of infection need further investigation. Additionally, multiple mycotoxins were detected and quantified in maize residues during the wheat cycle, raising the question of their ecological role and impact on the soil microbiota. [ABSTRACT FROM AUTHOR]

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Silage has been identified as a source of different microbial toxins, that may impair farm animal health and productivity as human health can also be compromised. In this sense, the aim of this study was to determine the impact of silage additives on the concentrations of deoxynivalenol (DON) and zearalenone (ZEN) mycotoxins and, eventually, to evaluate the hygienic quality of orchardgrass (Dactylis glomerata L.) silage based on the concentration of them compared to control silage. This study evaluated the influence of biological and chemical additives used in six different varieties of orchardgrass silage on DON and ZEN mycotoxin contents for the first time. The content of both fusariotoxins (DON and ZEN) in fresh matter and grass silage were below the threshold stipulated by the European Commission. The concentration of DON ranges from ~21.86 to 37.26 ng/kg, ~10.21 to 15 ng/kg, ~20.72 to 29.14 ng/kg; and ZEN range from ~3.42 to 7.87 ng/kg, ~3.85 to 8.62 ng/kg and ~2.15 to 5.08 ng/kg, in control, biological and chemical silages, respectively. In general, the biological additive was more efficient for preventing DON contamination, whereas the chemical additive was more efficient for preventing ZEN contamination in grass silage. In summary, the results obtained in this work demonstrate that biological and chemical additives can inhibit fungal growth and mycotoxin production on Dactylis glomerata L. silage and whose use could prevent animal and human diseases. [ABSTRACT FROM AUTHOR]

Background: Deoxynivalenol (DON) is a type B trichothecene mycotoxin that is commonly found in cereals and grains worldwide. The presence of this fungal secondary-metabolite raises public-health concerns at both the agriculture and food industry level. Recently, we have shown that DON has a negative impact on gut integrity, a feature also noticed for Campylobacter (C.) jejuni. We further demonstrated that DON increased the load of C. jejuni in the gut and inner organs. In contrast, feeding the less toxic DON metabolite deepoxy-deoxynivalenol (DOM-1) to broilers reduced the Campylobacter load in vivo. Consequently, it can be hypothesized that DON and DOM-1 have a direct effect on the growth profile of C. jejuni. The aim of the present study was to further resolve the nature of this interaction in vitro by co-incubation and RNA-sequencing. Results: The co-incubation of C. jejuni with DON resulted in significantly higher bacterial growth rates from 30 h of incubation onwards. On the contrary, the co-incubation of C. jejuni with DOM-1 reduced the CFU counts, indicating that this DON metabolite might contribute to reduce the burden of C. jejuni in birds, altogether confirming in vivo data. Furthermore, the transcriptomic profile of C. jejuni following incubation with either DON or DOM-1 differed. Co-incubation of C. jejuni with DON significantly increased the expression of multiple genes which are critical for Campylobacter growth, particularly members of the Flagella gene family, frr (ribosome-recycling factor), PBP2 futA-like (Fe3+ periplasmic binding family) and PotA (ATP-binding subunit). Flagella are responsible for motility, biofilm formation and host colonization, which may explain the high Campylobacter load in the gut of DON-fed broiler chickens. On the contrary, DOM-1 downregulated the Flagella gene family and upregulated ribosomal proteins. Conclusion: The results highlight the adaptive mechanisms involved in the transcriptional response of C. jejuni to DON and its metabolite DOM-1, based on the following effects: (a) ribosomal proteins; (b) flagellar proteins; (c) engagement of different metabolic pathways. The results provide insight into the response of an important intestinal microbial pathogen against DON and lead to a better understanding of the luminal or environmental acclimation mechanisms in chickens. [ABSTRACT FROM AUTHOR]

Avian coccidiosis, a common disease caused by Eimeria species, results in significant losses in global poultry production. Mycotoxins are low-molecularweight natural products (i.e., small molecules) produced as secondary metabolites by filamentous fungi and they have the potential to economically and significantly affect global poultry production. Little is known about the relationship between mycotoxins and avian coccidiosis, although they often co-occur in the field. This comprehensive review examines the intricate relationship between mycotoxins and avian coccidiosis, in particular how mycotoxins, including aflatoxins, ochratoxins, trichothecenes as well as Fusarium mycotoxins, compromise the health of the poultry flock and open the door to Eimeria parasites in the gut. In addition, this review sheds light on the immunosuppressive effects of mycotoxins, their disruption of cellular signaling pathways, and the consequent exacerbation of coccidiosis infections. The mechanisms of mycotoxin toxicity are also reviewed, emphasizing direct damage to intestinal epithelial cells, impaired nutrient absorption, inflammation, oxidative stress, and changes in the gut microbiota. Finally, the consequences for the prevention and treatment of coccidiosis when mycotoxins are present in the feed are discussed. This review emphasizes the need for effective management strategies to mitigate the combined risks of mycotoxins and coccidiosis and highlights the complexity of diagnosing and controlling these interrelated problems in poultry. The review advocates a holistic approach that includes strict feed management, disease prevention measures and regular monitoring to maintain the health and productivity of poultry against these significant challenges. [ABSTRACT FROM AUTHOR]

The experiment aimed to explore the effects of different water contents and storage time on the content of mycotoxins in the mushroom bran of Flammulina velutifolia. This study set up 13%, 30%, 48%, 62%, and 76% moisture gradients of Flammulina velutifolia bran, and measured the content of aflatoxin B1 (AFB1), zearalenone (ZEN), ochratoxin A (OTA), and T-2 toxin (T-2) in Flammulina velutifolia bran, respectively. The dynamic changes in the content of Flammulina velutifolia bran naturally stored for 1~7 days were recorded, and the correlation between the content of each Flammulina velutifolia toxin and storage time was analyzed. Combined with the requirements of the Feed Hygiene Standard for the content of Flammulina velutifolia bran in feed raw materials, it was determined whether the Flammulina velutifolia bran exceeded the limit of mycotoxins. The results showed that at a moisture content of 13%, the contents of AFB1, ZEN, OTA, and T-2 in the mushroom bran of Flammulina velutifolia showed no significant differences within 1~7 days after storage (P>0.05), and none of them exceeded the standard threshold. At 30%, 48%, 62%, and 76% moisture content, the content of AFB1, ZEN, OTA, and T-2 in Flammulina velutifolia bran showed an increasing trend with prolonged storage time. At a moisture content of 30%, the content of AFB1, ZEN, OTA, and T-2 in the Flammulina velutifolia bran did not exceed the standard threshold. At 48%, 62%, and 76% moisture content, the content of AFB1, ZEN, OTA, and T-2 in the Flammulina velutifolia bran did not exceed the standard within 1~2 days after storage. At a moisture content of 62%, the content of AFB1 in the Flammulina velutifolia bran exceeded the standard threshold after the 3rd day, and all three contents exceeded the standard threshold on the 5th day. At 62% and 76% moisture content, both ZEN and OTA in the Flammulina velutifolia bran exceeded the standard threshold on the 7th, with T-2 content not exceeding the standard threshold. At low moisture content (13%), there was no significant correlation between fungal toxin content and storage time (P>0.05). Under medium to high moisture content (48% and 76%), there was a significant positive correlation between fungal toxin content and storage time (P<0.05), and there was a significant positive correlation between different fungal toxins (P<0.05). The study indicates that when safely storing Flammulina velutifolia bran, it is necessary to control the moisture content and storage time. When the moisture content of Flammulina velutifolia bran is below 13% or high moisture Flammulina velutifolia bran is used within two days of leaving the factory, it can ensure that the content of mycotoxins does not exceed the standard. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEN) is a fungal toxin produced by Fusarium that widely occurs in various types of grains and feed. It has strong reproductive toxicity and a wide range of distribution, which will impact food and feed safety. In this study, Bacillus subtilis ZENL09, which is capable of transforming ZEN was investigated. By optimizing the fermentation conditions, the supernatant of Bacillus subtilis ZENL09 fermentation can achieve about 88% degradation rate of ZEN after reacting with ZEN for 12 h. The degradation product was collected by high-performance liquid chromatography, followed by LC-Q-TOF and MS/MS analysis to identify the changes in molecular mass and compound structure. The possible structure of the degradation product was also tentatively determined. Further evaluation of the degradation products' cytotoxicity was conducted using HepG2 cells, and the results indicated a significant reduction in toxicity compared to ZEN. Detoxification experiments on corn steep liquor showed that the fermentation supernatant of Bacillus subtilis degraded 87% of ZEN within 12 h, which further validates its potential application for detoxification in food and agricultural products. [ABSTRACT FROM AUTHOR]

Simple Summary: Aflatoxin B1 (AFB1) is a potent mycotoxin that can severely harm broiler health and performance, leading to reduced growth rates, compromised liver function, and weakened immune function. The study explored the effectiveness of two dietary supplements, hydrated sodium calcium aluminosilicates (HSCASs), and discarded date pits (DDPs) in reducing the harmful effects of AFB1 on broiler chickens. The research involved feeding 240 chickens with different diets: a control diet, a control diet with AFB1, and an AFB1-contaminated diet with either HSCAS or DDP. The results showed that including HSCAS or DDP in the diet improved the chickens' body weight, feed efficiency, and overall health. These improvements were evident through better liver and kidney function, higher protein and glucose levels in the blood, and enhanced digestive efficiency. Remarkably, DDP supplementation also reduced liver size and improved total antioxidant capacity levels, while HSCAS increased protein digestibility. The study concluded that both HSCAS and DDP are effective in reducing the negative impacts of AFB1 on broiler chickens, suggesting they could be valuable natural feed additives in poultry farming to ensure healthier birds and safer food products for consumers. The research aimed to evaluate how effective hydrated sodium calcium aluminosilicates (HSCASs) and discarded date pits (DDPs) are as dietary adsorbents for aflatoxin B1 (AFB1) in enhancing the performance and health of broiler chickens aged 16 to 30 days. A total of 240 Ross 308 straight-run broilers were randomly allocated into four dietary groups, each with 10 replicates: a control diet, a control diet with 1000 ppb AFB1, an AFB1-contaminated diet with 0.5% HSCAS, and an AFB1-contaminated diet with 4% DDP. Incorporating HSCASs or DDPs into the AFB1-contaminated diet resulted in significant improvements across various parameters, involving increased body weight, improved feed conversion ratio, higher dressing percentage, decreased relative weights of kidney and spleen, elevated serum levels of total protein, globulin, and glucose, reduced serum alanine aminotransferase activity, and heightened hepatic protein concentration and glutathione peroxidase activity, along with diminished hepatic malondialdehyde content and glutamic oxaloacetic transaminase activity. Moreover, both supplements led to increased ileal villus height and surface area, enhanced apparent nitrogen-corrected metabolizable energy digestibility, and decreased AFB1 residues in the liver and kidney. Moreover, the dietary inclusion of DDPs significantly decreased relative liver weight, raised serum albumin concentration, lowered serum alkaline phosphatase activity, enhanced hepatic total antioxidant capacity level, and augmented ileal villus width. Conversely, the dietary addition of HSCASs significantly heightened apparent crude protein digestibility. In conclusion, the inclusion of HSCASs and DDPs in AFB1-contaminated diets can mitigate the toxic effects of AFB1 on broiler chickens, with DDPs exhibiting additional advantages in optimizing liver function and gut morphology. [ABSTRACT FROM AUTHOR]

The present study aimed to find whether low doses of mixed mycotoxins would affect egg quality in laying hens, and to explore the oxidative stress induced liver damage through endoplasmic reticulum during summer stress. A total of 96 Jinghong laying hens, 36 wks of age, were divided into four treatments, with eight repetitions per treatment and three hens per repetition. All the hens were raised in summer (average temperature: 31.3 ± 0.5℃; average humidity: 85.5 ± 0.2%) for 28d. One treatment was fed a basal diet as control (CON), and the other three treatments were fed the same diets containing 3.0 mg/kg deoxynivalenol (DON), 0.5 mg/kg T-2 toxin (T-2), and 1.5 mg/kg DON + 0.25 mg/kg T-2 toxin (Mix). Albumen height and Haugh unit were decreased (P < 0.05) in the Mix group on day 14 and 28. The activity of total antioxidant capacity, glutathione peroxidase, catalase, and superoxide dismutase were decreased (P < 0.05) in the DON, T-2, and Mix groups. The alkaline phosphatase level in DON, T-2, and Mix groups was significantly increased (P < 0.05). The level of interleukin-1β, interferon-γ, and tumor necrosis factor-α in the Mix group were higher (P < 0.05) than CON, DON, and T-2 groups. Mix group upregulated the mRNA expressions of protein kinase RNA-like ER kinase, activating transcription factor4, IL-1β, nuclear factor-κ-gene binding, and nuclear respiratory factor 2 in the liver (P < 0.05). The results showed that low doses of DON and T-2 toxin could cause oxidative stress in the liver, but DON and T-2 toxin have a cumulative effect on virulence, which can reduce egg quality and cause endoplasmic reticulum stress in the liver. [ABSTRACT FROM AUTHOR]

The presence of mycotoxins and other toxic metabolites in hops (Humulus lupulus L.) was assessed for the first time. In total, 62 hop samples were sampled in craft breweries, and analyzed by a multi-toxin LS-MS/MS method. The study collected samples from craft breweries in all of the Croatian counties and statistically compared the results. Based on previous reports on Alternaria spp. and Fusarium spp. contamination of hops, the study confirmed the contamination of hops with these toxins. Alternaria toxins, particularly tenuazonic acid, were found in all tested samples, while Fusarium toxins, including deoxynivalenol, were present in 98% of samples. However, no Aspergillus or Penicillium metabolites were detected, indicating proper storage conditions. In addition to the Alternaria and Fusarium toxins, abscisic acid, a drought stress indicator in hops, was also detected, as well as several unspecific metabolites. The findings suggest the need for monitoring, risk assessment, and potential regulation of Alternaria and Fusarium toxins in hops to ensure the safety of hop usage in the brewing and pharmaceutical industries. Also, four local wild varieties were tested, with similar results to the commercial varieties for toxin contamination, but the statistically significant regional differences in toxin occurrence highlight the importance and need for targeted monitoring. [ABSTRACT FROM AUTHOR]

Using alternative ingredients or low-quality grain grades to reduce feeding costs for pig diets can introduce mycotoxins such as deoxynivalenol (DON) into feed, which is known to induce anorexia, inflammation, and oxidative stress. Adding vitamin 25(OH)D3 or vitamins E and C to the feed could increase piglets' immune system to alleviate the effects of DON. This study used 54 pigs (7.8 ± 0.14 kg) in 27 pens (2 pigs/pen) with a vitamin 25(OH)D3 or vitamin E-C supplementation, or their combination, in DON-contaminated (5.1 mg/kg) feed ingredients over 21 days followed by a lipopolysaccharide (LPS) challenge (20 µg/kg BW) 3 h prior to euthanasia for 1 piglet per pen. DON contamination induced anorexia, which reduced piglet growth. DON also induced immunomodulation, oxidative stress, and downregulated vitamin D status. The vitamin E and C supplementation and the combination of vitamins E, C, and 25(OH)D3 provided protection against DON contamination by not only decreasing blood and liver oxidative stress markers, but also by increasing antioxidant enzymes and tocopherol levels in blood, indicating improved antioxidant defense mechanisms. The combination of vitamins also restored the vitamin D status. After LPS challenge, DON contamination decreased intestinal and liver antioxidant statuses and increased inflammation markers. The addition of vitamins E and C to DON-contaminated feed reduced markers of inflammation and improved the antioxidant status after the LPS immune stimulation. The combination of all these vitamins also reduced the oxidative stress markers and the inflammation in the intestine and mesenteric lymph nodes, suggesting an anti-inflammatory effect. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) is a prevalent toxin causing severe liver damage through hepatocellular oxidative stress. However, the underlying mechanisms and effective therapeutic approaches remain unknown. Here, the unique role of the xenobiotic metabolism factor pregnane X receptor (PXR) in mediating DON‐induced hepatocellular oxidative stress is investigated. Treatment with the PXR agonist 3‐indole‐propionic acid (IPA) alleviates DON‐induced oxidative stress and liver injury both in vitro and in vivo. Mechanistically, it is discovered for the first time that PXR agonist IPA directly transactivates the m6A demethylase FTO expression, leading to site‐specific demethylation and decreased abundance of YTHDC1‐bound Malat1 lncRNA at single‐nucleotide resolution. The diminished m6A modification of Malat1 lncRNA reduces its stability and augments antioxidant pathways governed by NRF2, consequently mitigating DON‐induced liver injury. Furthermore, Malat1 knockout mice exhibit decreased DON‐induced liver injury, emphasizing the role of Malat1 lncRNA in oxidative stress. Collectively, the findings establish that PXR‐mediated m6A‐dependent Malat1 lncRNA expression determines hepatocyte oxidative stress via m6A demethylase FTO, providing valuable insights into the potential mechanisms underlying DON‐induced liver injury and offers potential therapeutic strategies for its treatment. [ABSTRACT FROM AUTHOR]

Ochratoxin A (OTA) is a mycotoxin commonly found in improperly stored grains, posing significant risks to food safety. Accurate detection of this mycotoxin in food matrices is crucial for ensuring consumer protection. In this study, we used in silico methods to design new peptides with high affinity and selectivity for recognizing OTA, starting crystal structures of OTA-protein complexes, which served to identify short key peptides within the interaction regions. Through deliberate point mutations, the stability and affinity of these peptides towards OTA were enhanced. To assess their suitability as capture probes for OTA sensing, we investigated the interaction of the selected peptides with OTA in a saline aqueous solution to mimic wet-lab experiments. Molecular docking, molecular dynamics simulations were used to evaluate the affinity and stability of the peptide-OTA complexes. Various parameters, including conformational changes, Molecular Mechanics Poisson–Boltzmann Surface Area energetics, the number of contacts, and the number of hydrogen bonds were analyzed to identify the most promising candidates. Among them, the peptide CWC11 has the highest affinity to OTA. Next, CWC11 peptide was used to functionalize the surfaces of two different carbonaceous nanomaterials: reduced graphene oxide (rGO) and graphene oxide (GO). The performance of the peptide-functionalized surfaces in presence of OTA was evaluated, revealing that GO exhibited superior performance compared to rGO. This study provides valuable insights into the design OTA-binding peptides and their use for the functionalization of carbonaceous surfaces, emphasizing the importance of surface selection to tune the efficiency and sensitivity of the detection. [ABSTRACT FROM AUTHOR]

The fungal genus Fusarium contains many important plant pathogens as well as endophytes of wild and crop plants. Globally, Fusarium toxins in food crops are considered one of the greatest food safety concerns. Their occurrence has become more pronounced in Africa in recent times. Among the major Fusarium mycotoxins with food and feed safety concerns, zearalenone is frequently detected in finished feeds and cereals in Africa. However, the impact of indigenous agricultural practices (pre‐ and postharvest factors) and food processing techniques on the prevalence rate of Fusarium species and zearalenone occurrence in food and feed have not been collated and documented systematically. This review studies and analyzes recent reports on zearalenone contamination in maize and other cereal products from Africa, including its fungi producers, agronomic and climate variables impacting their occurrences, preventive measures, removal/decontamination methods, and legislations regulating their limits. Reports from relevant studies demonstrated a high prevalence of F. verticillioides and F. graminearum as Africa's main producers of zearalenone. Elevated CO2 concentration and high precipitation may carry along an increased risk of zearalenone contamination in maize. African indigenous processing methods may contribute to reduced ZEA levels in agricultural products and foods. Most African countries do not know their zearalenone status in the food supply chain and they have limited regulations that control its occurrence. [ABSTRACT FROM AUTHOR]

The massive import of uncontrolled technical grain from the East into the European Community poses a risk to public health when it ends up in the mills to be used as flour for food purposes instead of being burnt (biofuel). In fungal infections of wheat, the most dangerous species belong to the genus Fusarium. F. poae is a pathogen that is most commonly isolated from cereals worldwide and causes various types of diseases in animals and humans due to the numerous toxins it produces. The manuscript reports an attempt to distinguish between four species of Fusarium, F. avanceum, F. langsethiae, F. poae, and F. sporotrichioides, in wheat grains by measuring the volatiles emitted. The patterns obtained from the signals captured by the electronic nose PEN3 were used to build the Random Forests classification model. The recall and precision of the classification performance for F. poae reached 91 and 87%, respectively. The overall classification accuracy reached 70%. Gas chromatography coupled with mass spectrometry (GC–MS) was used to analyze the chemical composition of the emitted volatiles. The patterns found in the GC–MS results allowed an explanation of the main patterns observed when analyzing the electronic nose data. The mycotoxins produced by the Fusarium species analyzed were detected. The results of the reported experiment confirm the potential of the electronic nose as a technology that can be useful for screening the condition of the grain and distinguishing between different pathogenic infestations. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) is a common mycotoxin that contaminates cereals. Therefore, the development of sensitive and efficient detection methods for DON is essential to guarantee food safety and human health. In this study, an enzyme cascade amplification-based immunoassay (ECAIA) using a dual-functional alkaline phosphatase-linked single-chain fragment variable fusion tracer (scFv-ALP) and MnO2 nanosheets was established for DON detection. The scFv-ALP effectively catalyzes the hydrolysis of ascorbyl-2-phosphate (AAP) to produce ascorbic acid (AA). This AA subsequently interacts with MnO2 nanosheets to initiate a redox reaction that results in the loss of oxidizing properties of MnO2. In the absence of ALP, MnO2 nanosheets can oxidize 3,3′,5,5′-tetramethylbenzidine (TMB) to produce the blue oxidized product of TMB, which exhibits a signal at a wavelength of 650 nm for quantitative analysis. After optimization, the ECAIA had a limit of detection of 0.45 ng/mL and a linear range of 1.2–35.41 ng/mL. The ECAIA exhibited good accuracy in recovery experiments and high selectivity for DON. Moreover, the detection results of the actual corn samples correlated well with those from high-performance liquid chromatography. Overall, the proposed ECAIA based on the scFv-ALP and MnO2 nanosheets was demonstrated as a reliable tool for the detection of DON in corn samples. [ABSTRACT FROM AUTHOR]

Pig farming in India faces challenges in meeting the growing demand for pork, necessitating improvements in nutrient utilization, feed conversion, and growth rates in pigs. Mycotoxins are naturally occurring fungal toxins that can contaminate pig feed at various stages of production, leading to adverse health effects in pigs. The study was designed to assess the efficacy of supplementation of herbal product in mitigating the negative effects of low levels of mycotoxins in the diet and economics involved in the cost of production. The 18 weaned large white Yorkshire piglets of four weeks age with body weight 6-8 kg were randomly divided into three groups (T0, T1 and T2) comprising of six piglets in each group. All the animals were reared for a period of four months and were fed with mycotoxin levels at maximum permissible levels. The animals in T0 were not given herbal supplementation. The animals in T1 were supplemented with herbal supplementation and T2 were supplemented with Commercial mycotoxin binder. The experimental animals were observed fortnightly interval for body weight gain, feed conversion ratio (FCR), carcass characters and hemato-biochemical parameters. At 8th fortnight interval; body weight was significantly high in T1 and T2 and significantly low in T0 groups. The overall FCR of four months showed significant differences among different groups and was significantly high in T0, and low in T1 group. At the end of 5th month age, hot carcass weight, dressing percentage, carcass length, meat percent were significantly high in T1 and T2 and low in T0 group. It has been inferred that the use of herbomineral mixture and toxin binders in the feed is very advantageous in reducing adverse effect of mycotoxins on piglet performance. The herbomineral mixture was found effective similar to commercial toxin binder. [ABSTRACT FROM AUTHOR]

Fusarium head blight (FHB) is a destructive disease caused by several species of Fusarium, such as Fusarium graminearum and F. asiaticum. FHB affects cereal crops, including wheat, barley, and rice, worldwide. Fusarium-infected kernels not only cause reduced yields but also cause quality loss by producing mycotoxins, such as trichothecenes and zearalenone, which are toxic to animals and humans. For decades, chemical fungicides have been used to control FHB because of their convenience and high control efficacy. However, the prolonged use of chemical fungicides has caused adverse effects, including the emergence of drug resistance to pathogens and environmental pollution. Biological control is considered one of the most promising alternatives to chemicals and can be used for integrated management of FHB due to the rare possibility of environment pollution and reduced health risks. In this study, Bacillus velezensis JCK-7158 isolated from rice was selected as an ecofriendly alternative to chemical fungicides for the management of FHB. JCK-7158 produced the extracellular enzymes protease, chitinase, gelatinase, and cellulase; the plant growth hormone indole-3-acetic acid; and the 2,3-butanediol precursor acetoin. Moreover, JCK-7158 exhibited broad antagonistic activity against various phytopathogenic fungi and produced iturin A, surfactin, and volatile substances as active antifungal compounds. It also enhanced the expression of PR1, a known induced resistance marker gene, in transgenic Arabidopsis plants expressing ß-glucuronidase (GUS) fused with the PR1 promoter. Under greenhouse conditions, treatments with the culture broth and suspension concentrate formulation of JCK-7158 at a 1,000-fold dilution inhibited the development of FHB by 50 and 66%, respectively. In a field experiment, treatment with the suspension concentrate formulation of JCK-7158 at a 1,000-fold dilution effectively controlled the development of FHB with a control value of 55% and reduced the production of the mycotoxin nivalenol by 40%. Interestingly, treatment with JCK-7158 enhanced the expression of plant defense-related genes in salicylic acid, jasmonic acid, ethylene, and reactive oxygen species (ROS) signaling pathways before and after FHB pathogen inoculation. Taken together, our findings support that JCK-7158 has the potential to serve as a new biocontrol agent for the management of FHB. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the effectiveness of nine different biological compounds to reduce mycotoxins concentrations. The hypothesis of this study was that a static in vitro gastrointestinal tract model, as an initial screening tool, can be used to simulate the efficacy of Geotrichum fermentans, Rhodotorula rubra, Kluyveromyce marxiamus yeast cell walls and their polysaccharides, red and white clay minerals, and walnuts nutshells claiming to detoxify AFB1, ZEA, DON, and T-2 toxin mycotoxins. Mycotoxin concentrations were analyzed using high-performance liquid chromatography (HPLC) with fluorescent (FLD) and ultraviolet detectors (UV). The greatest effects on reducing mycotoxin concentrations were determined as follows: for AFB1, inserted G. fermentans cell wall polysaccharides and walnut nutshells; for ZEA, inserted R. rubra and G. fermentans cell walls and red clay minerals; for DON, R. rubra cell wall polysaccharides and red clay minerals; and for T-2 toxin, R. rubra cell walls, K. marxianus, and G. fermentans cell wall polysaccharides and walnut nutshells. The present study indicated that selected mycotoxin-detoxifying biological compounds can be used to decrease mycotoxin concentrations. [ABSTRACT FROM AUTHOR]

Endometrial cancer is one of the most common cancer types among women. Many factors can contribute to the development of this disease, including environmental factors and, thus, eating habits. Our study aims to determine the levels of various mycotoxins and their metabolites in the blood serum and endometrial tissue samples of participants with previously proven endometrial cancer and to find possible contributions to cancer development. In the cohort clinical trial, 52 participants aged between 44 and 86 were studied. The participants were divided into two groups: patients or matched controls. All patients had previously histologically diagnosed endometrial cancer. The cancer patients were divided into low-grade endometrioid and low- plus high-grade endometrioid groups. Controls had no history of endometrial malignancy or premalignancy. Blood serum and endometrial tissue samples were obtained from all study patients. We compared the concentrations of total Aflatoxins (Afs), Deoxynivalenol (DON), Ochratoxin-A (OTA), T2-toxin and HT2 toxin (T2/HT2 toxin), Zearalenone (ZEN), alpha-Zearalenol (α-ZOL), and Fumonisin B1 (FB1) in the serum and endometrium between the different study groups. As a result, we can see a significant correlation between the higher levels of Afs and zearalenone and the presence of endometrial cancer. In the case of Afs, DON, OTA, T2/HT2 toxins, ZEN, and alpha-ZOL, we measured higher endometrial concentrations than in serum. Considering the effect of mycotoxins and eating habits on cancer development, our results might lead to further research exploring the relationship between certain mycotoxins and endometrium cancer. [ABSTRACT FROM AUTHOR]

Fusarium pathogens and their mycotoxins are considered as the main threats to cereal production and food safety worldwide. However, due to the constant discovery of new Fusarium species especially along with mycotoxin production profile differentiation in certain species, efforts on their species composition, geographical distribution, and chemotype proportion are urgently required. In the Fusarium goolgardi species, two distinct trichothecene (TB) genotype populations have recently been identified. Previous studies have shown that the structural variance of TBs biosynthesized by the two genotypes is attributed to the Tri1 gene. Polymorphisms of Tri1 gene from type A TB-producers were investigated in different Fusarium species in the current study. According to these DNA sequence variations identified in Tri1 gene sequences, a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based diagnostic approach for the differentiation of T-2 and 4,15-diacetoxyscirpenol (DAS) genotypes in F. goolgardi was successfully developed. The PCR-RFLP assay will facilitate the studies on geographic distribution, frequency and other aspects of the two genotypes within F. goolgardi species. [ABSTRACT FROM AUTHOR]

The market demand for herbal dietary supplements is rapidly growing and such products are becoming more common and accessible to consumers. However, the knowledge about their safety remains incomplete. Herbal dietary supplements are one of the food groups that can contribute significantly to human health concerns arising from chronic exposure to pyrrolizidine alkaloids and mycotoxins. This study aimed to simultaneously determine 79 natural contaminants, including mycotoxins, as well as pyrrolizidine and tropane alkaloids in herbal dietary supplements in one analytical run. Exposure assessment and human health risks were assessed for all compounds included in this study. The total concentration of naturally occurring contaminants in herbal dietary supplements reached 5.3 mg kg−1 and the most frequently detected mycotoxins were tentoxin and alternariol monomethyl ether. The latter was detected with the highest frequency, reaching concentrations up to 2.5 mg kg−1. The obtained results indicate a potential risk to public health related to herbal dietary supplement consumption. [ABSTRACT FROM AUTHOR]

Hippocampal neurons maintain the ability of proliferation throughout life to support neurogenesis. Deoxynivalenol (DON) is a mycotoxin that exhibits brain toxicity, yet whether and how DON affects hippocampal neurogenesis remains unknown. Here, we use mouse hippocampal neuron cells (HT-22) as a model to illustrate the effects of DON on neuron proliferation and to explore underlying mechanisms. DON exposure significantly inhibits the proliferation of HT-22 cells, which is associated with an up-regulation of cell cycle inhibitor p21 at both mRNA and protein levels. Global and site-specific m6A methylation levels on the 3'UTR of p21 mRNA are significantly increased in response to DON treatment, whereas inhibition of m6A hypermethylation significantly alleviates DON-induced cell cycle arrest. Further mechanistic studies indicate that the m6A readers YTHDF1 and IGF2BP1 are responsible for m6A-mediated increase in p21 mRNA stability. Meanwhile, 3'UTR of E3 ubiquitin ligase TRIM21 mRNA is also m6A hypermethylated, and another m6A reader YTHDF2 binds to the m6A sites, leading to decreased TRIM21 mRNA stability. Consequently, TRIM21 suppression impairs ubiquitin-mediated p21 protein degradation. Taken together, m6A-mediated upregulation of p21, at both post-transcriptional and post-translational levels, contributes to DON-induced inhibition of hippocampal neuron proliferation. These results may provide new insights for epigenetic therapy of neurodegenerative diseases. DON inhibits the proliferation of HT-22 cells. RNA m6A hypermethylation on the transcript of p21 enhances the mRNA stability in a YTHDF1- and IGF2BP1-dependent manner, which leads to the upregulation of p21. RNA m6A hypermethylation on the transcript of TRIM21 decreases the mRNA stability in a YTHDF2-dependent manner, which contributes to prevent p21 ubiquitin-mediated degradation. High expression of p21 contributes to inhibit cell proliferation. [ABSTRACT FROM AUTHOR]

Simple Summary: Early embryonic mortality is one of the main causes of poor reproductive health and is influenced by many factors, including mycotoxins and their metabolites in feed. When several mycotoxins are present in feed, they can have different effects and cause significant animal health problems; however, their synergistic effects have not yet been explored. We found that the blastocyst rate was affected by acetonitrile as a vehicle for the mycotoxin. When the desired mycotoxin concentrations were diluted with the lowest possible amount of acetonitrile, we found no statistical difference between blastocyst rates, although there was a trend suggesting that the combination caused poorer blastocyst development. However, a less invasive vehicle should be used to see if the combination of mycotoxins can affect blastocyst rates. The quality of animal feed is increasingly affected by weather conditions, high humidity, and damage to grains, which have led to various mycotoxin-producing moulds. The aim of this study was to determine the effects of the combination of deepoxy-deoxynivalenol and beta-zearalenol on the development of preimplantation bovine embryos, the extent to which the presence of both mycotoxin metabolites affects the development of in vitro cultured bovine embryos, or whether the effect of both toxins enhances embryotoxicity. Ovaries were transported from the abattoir to the laboratory and, after maturation and fertilisation, zygotes were placed in an embryo culture medium (IVC) with different mycotoxin metabolite concentrations diluted in acetonitrile. It was found that the blastocyst rate of cleaved embryos was affected by 1 μL acetonitrile in 400 μL medium (0.25%) compared to the group without acetonitrile. For this reason, it was decided to use acetonitrile as a control group, and the desired mycotoxin metabolite concentrations were diluted in the lowest possible amount of acetonitrile (0.5 μL) that could be accurately added to the study groups. There was no statistical difference when the higher mycotoxin metabolite concentrations were added. [ABSTRACT FROM AUTHOR]